Abstract [en]

Background and Objectives:The cytolethal distending toxin (Cdt) is a highly conserved exotoxin that are produced by a number of Gram negative bacteria, including Aggregatibacter actinomycetemcomitans, and affects mammalian cells by inhibiting cell division and causing apoptosis. A complete cdt-operon is present in the majority of A. actinomycetemcomitans, but the proportion of isolates that lack cdt-encoding genes (A, B and C) varies according to the population studied. The objectives of this study were to examine serotype, Cdt-genotype, and Cdt-activity in isolates of A. actinomycetemcomitans collected from an adolescent West African population and to examine the association between the carrier status of A. actinomycetemcomitans and the progression of attachment loss (AL).

Material and Methods: A total of 249 A. actinomycetemcomitans isolates from 200 Ghanaian adolescents were examined for serotype and cdt-genotype by PCR. The activity of the Cdt-toxin was examined by DNA-staining of exposed cultured cells and documented with flow cytometry. The periodontal status of the participants was examined at baseline and at a two-year follow-up.

Results: Presence of all three cdt-encoding genes was detected in 79% of the examined A. actinomycetemcomitans isolates. All these isolates showed a substantial Cdt-activity. The two different cdt-genotypes (with and without presence of all three cdt-encoding genes) showed a serotype-dependent distribution pattern. Presence of A. actinomycetemcomitans was significantly associated with progression of AL (OR = 5.126; 95% CI = [2.994 - 8.779], p < 0.001).

Conclusion: A. actinomycetemcomitans isolated from the Ghanaian adolescents showed a distribution of serotype and cdt-genotype in line with results based on other previously studied populations. Presence of A. actinomycetemcomitans was significantly associated with disease progression, in particular the b serotype, whereas the association with disease progression was not particularly related to cdt-genotype, and Cdt-activity.

Höglund Åberg, Carola

Umeå University, Faculty of Medicine, Department of Odontology.

2013 (English)Doctoral thesis, comprehensive summary (Other academic)

Abstract [en]

Aggregatibacter actinomycetemcomitans is an oral bacterium that is mainly associated with aggressive forms of periodontitis, which most often starts at an early age. Amongst the virulence factors of A. actinomycetemcomitans, two exotoxins, the leukotoxin (LtxA) and the cytolethal distending toxin (Cdt), are suggested to play an important role in the pathogenicity of aggressive periodontitis. There is also a genetic diversity of the different strains of A. actinomycetemcomitans, and a variation in the ability of different strains to express and release exotoxins has been suggested. Of the different genotypes of A. actinomycetemcomitans, the highly leukotoxic JP2 genotype, which is prevalent in individuals of African origin, seems to be the genotype that is most strongly associated with localized aggressive periodontitis.

This thesis is built upon studies of a West African adolescent population. The aim was to study the virulence characteristics of A. actinomycetemcomitans genotypes with a specific focus on the LtxA and the Cdt in relation to the progression of attachment loss (AL). The specific aim was first, to investigate cross-sectionally the presence of the JP2 and non-JP2 genotypes of A. actinomycetemcomitans in relation to the prevalence of AL and then prospectively to assess the progression of AL in a Ghanaian adolescent population. Second, in clinical isolates of A. actinomycetemcomitans obtained from the participants of the study, the serotypes and the virulence characteristics related to the two exotoxins were studied and associated with the progression of AL at the individual level.

In Paper I, based on the study population consisting of 500 adolescents (mean age 13.2 years; SD ±1.5), it was shown that the overall carrier rate of A. actinomycetemcomitans was high (54.4%) and that the presence of this bacterium was associated with AL ≥ 3 mm. The JP2 genotype was prevalent (8.8%) in this population. In Paper II, 397 (79.4%) of the study participants were periodontally examined again at a 2-year follow-up. The presence of the JP2 genotype of A. actinomycetemcomitans in subgingival plaquewas strongly associated with the progression of AL. This study also provided support for an enhanced estimated risk (odds ratio, OR=3.4), though less pronounced, for the progression of AL in individuals positive for the non-JP2 genotypes of A.actinomycetemcomitans.

In Paper III, all three cdt genes (a, b and c) were detected in 79% of the examined A. actinomycetemcomitans isolates, all of which expressed an active toxin. The distribution of the cdt genes showed a serotype-dependent pattern. In particular, the presence of the b serotypes (both JP2 and non-JP2 genotypes) was associated with the disease progression, whereas the expression of Cdt was not particularly related to the disease progression. In Paper IV, it was shown that the presence of of A. actinomycetemcomitans isolates with high leukotoxicity, also those of the non-JP2 genotypes of A. actinomycetemcomitans, were associated with an increased risk of the progression of AL in relation to the reference group. The main proportion of the serotype b isolates was distributed in the category of highly leukotoxic isolates. The analyses of the non-JP2 genotypes of serotype b indicated a diversity linked to the level of leukotoxicity.

In conclusion, A. actinomycetemcomitans in general was associated with the progression of AL. Individuals with an increased risk of developing progression of AL mainly harboured isolates of A. actinomycetemcomitans with a high leukotoxicity, which suggests that the LtxA is an important virulence factor. Of the two exotoxins, the pathogenic potential was mainly associated with the LtxA, while the role of the Cdt is unclear.